The present invention concerns palladium-substituted bacteriochlorophyll derivatives, processes and intermediates for their preparation and pharmaceutical compositions comprising the same as well as their use in the field of in vivo photodynamic therapy and diagnosis and in vitro photodynamic killing of viruses and microorganisms.
BChl=bacteriochlorophyll a (Mg-containing 7,8,17,18-tetrahydroporphyrin having a phytyl or geranylgeranyl group at position 173, a COOCH3 group at position 132, an H atom at position 132, an acetyl group at position 3 and an ethyl group at position 8).
BChlide=bacteriochlorophyllide a (the C-172-free carboxylic acid derived from BChl a).
BPhe=bacteriopheophytin a (BChl in which the central Mg atom is replaced by two H atoms).
BPheid=bacteriopheophorbide a (the C-172-free carboxylic acid derived from BPhe).
Pd-BPheid=Pd-bacteriopheophorbide a (the C-172-free carboxylic acid derived from BPhe having a central Pd atom, a COOCH3 group at position 132, an H atom at position 132, an acetyl group at position 3 and an ethyl group at position 8).
IUPAC numbering of the bacteriochlorophyll derivatives is used throughout the specification. Using this nomenclature, the natural bacteriochlorophylls carry two carboxylic acid esters at positions 132 and 172, however they are esterifed at positions 133 and 173.
There has been an increasing interest in the utilization of photosensitizers for cancer therapy. According to this technique, known as photodynamic therapy (PDT), photosensitizers are applied for example to a tumor and the in situ photosentization produces compounds which intoxicate the malignant cells.
Photodynamic therapy using porphyrins and related compounds has, by now, a fairly long history. Early work, in the 1940s, demonstrated that porphyrin could be caused to fluoresce in irradiated tumor tissue. The porphyrins appeared to accumulate in these tissues, and were capable of absorbing light in situ, providing a means to detect the tumor by the location of the fluorescence. A widely used preparation in the early stages of photodynamic treatment both for detection and for therapy was a crude derivative of hematoporphyrin, also called hematoporphyrin derivative, HpD, or Lipson derivative prepared as described by Lipson and coworkers in J Natl Cancer Inst (1961) 26:1-8. Considerable work has been done using this preparation, and Dougherty and coworkers reported the use of this derivative in treatment of malignancy (Cancer Res (1978) 38:2628-2635; J Natl Cancer Inst (1979) 62:231-237).
Dougherty and coworkers prepared a more effective form of the hematoporphyrin derivative which comprises a portion of HpD having an aggregate weight greater than 10 kd. This form of the drug useful in photodynamic therapy is the subject of U.S. Pat. No. 4,649,151, is commercially available, and is in clinical trials.
The general principles of the use of light-absorbing compounds, especially those related to porphyrins, has been well established as a treatment for tumors when administered systematically. The differential ability of these preparations to destroy tumor, as opposed to normal tissue, is due to the homing effect of these preparations to the objectionable cells. (See, for example, Dougherty, T. J., et al., xe2x80x9cCancer: Principles and Practice of Oncologyxe2x80x9d (1982), V. T. de Vita, Jr., et al., eds. pp 1836-1844.). Efforts have been made to improve the homing ability by conjugating hematoporphyrin derivative to antibodies. (See, for example, Mew, D., et al., J Immunol.(1983) 130:1473-1477.). The mechanism of these drugs in killing cells seems to involve the formation of singlet oxygen upon irradiation (Weishaupt, K. R., et al., Cancer Research (1976) pp. 2326-2329).
The use of hematoporphyrin derivative or its active components in the treatment of skin diseases using topical administration has also been described in U.S. Pat. No. 4,753,958. In addition, the drugs have been used to sterilize biological samples containing infectious organisms such as bacteria and virus (Matthews, J. L., et al., Transfusion (1988): 81-83). Various other photosensitizing compounds have also been used for this purpose, as set forth, for example, in U.S. Pat. No. 4,727,027.
In general, the methods to use radiation sensitizers of a variety of structures to selectively impair the functioning of biological substrates both in vivo and in vitro are understood in the art. The compounds useful in these procedures must have a differential affinity for the target biological substrate to be impaired or destroyed and must be capable of absorbing light so that the irradiated drug becomes activated in a manner so as to have a deleterious effect on the adjacent compositions and materials.
Because it is always desirable to optimize the performance of therapeutics and diagnostics, variations on the porphyrin drugs traditionally used in treatment and diagnosis have been sought. A number of general classes of photosensitizers have been suggested including phthalocyanines, psoralen-related compounds, and multicyclic compounds with resonant systems in general. Most similar to the compounds disclosed herein are various pheophorbide derivatives whose use in photodynamic therapy has been described in EPO Application 220686 to Nihon Metaphysics Company; ethylene diamine derivatives of pheophorbide for this purpose described in Japanese Application J85/000981 to Tama Seikayaku, K.K., and Japanese Application J88/004805 which is directed to 10-Hydroxypheophorbide-a. In addition, Beems, E. M., et al., in Photochemistry and Photobiology (1987) 46:639-643 disclose the use as photosensitizers of two derivatives of bacteriochlorophyll-axe2x80x94bacteriochlorophyllin-a (also known as bacteriopheophorbide-a, which lacks the phytyl alcohol derivatized in bacteriochlorophyll-a) and bacteriochlorin-a (which lacks both the phytyl group and the Mg ion). These authors direct their attention to these derivatives as being advantageous on the grounds of enhanced water solubility as compared to bacteriochlorophyll-a.
EP 584552 and WO97/19081, both to Yeda Research and Development Co. Ltd., describe chlorophyll and bacteriochlorophyll derivatives and their use as PDT agents, and metaled bacteriochrophylls and their preparation by transmetalation of the corresponding Cd-BChl derivatives, respectively.
The problem remains to find suitable photosensitizers useful in photodynamic therapy and diagnosis which are optimal for particular targets and particular contexts. Thus, the invention provides an additional group of photosensitizing compounds which becomes part of the repertoire of candidates for use in specific therapeutic and diagnostic situations.
It has now been found, in accordance with the present invention, that the compounds of formula I, Ixe2x80x2 or Ixe2x80x3 below wherein A as defined below represents a substituent capable of allowing an efficient plasma transfer and cell membrane penetration, are useful as PDT agents and present the advantages of enhanced solubility, stability and/or efficiency, compared with the known compounds.
The invention thus concerns the compounds of formula I, Ixe2x80x2, or Ixe2x80x3
wherein
A represents
OH,
OR1,
xe2x80x94Oxe2x80x94(CH2)nxe2x80x94Y,
xe2x80x94Sxe2x80x94(CH2)nxe2x80x94Y,
xe2x80x94NHxe2x80x94(CH2)nxe2x80x94Y,
xe2x80x94Oxe2x80x94(CH2)2xe2x80x94OH,
xe2x80x94NHxe2x80x94(CH2)nxe2x80x94+No,Xxe2x88x92,
xe2x80x94NHxe2x80x94(CH2)2xe2x80x94NHxe2x80x94BOC or
xe2x80x94Nxe2x80x94(CH2xe2x80x94CHxe2x95x90CH2)2 
wherein
R1 represents
Na+, K+, (Ca2+)0.5, (Mg2+)0.5, Li+, NH4+
+NH3xe2x80x94C(CH2OH)3, +NH3xe2x80x94CH2xe2x80x94(CHOH)4xe2x80x94CH2OH,
+NH2(CH3)xe2x80x94CH2xe2x80x94(CHOH)4xe2x80x94CH2OH or
+N(Cnxe2x80x2H2nxe2x80x2+1)4;
R2 represents H, OH or COOR4, wherein R4 is C1-C12 alkyl or C3-C12 cycloalkyl;
R3 represents H, OH or C1-C12 alkyl or alkoxy;
n is 1, 2, 3, 4, 5 or 6,
Y is xe2x80x94NRxe2x80x21Rxe2x80x22 or xe2x80x94+NRxe2x80x21Rxe2x80x22Rxe2x80x23, Xxe2x88x92 wherein Rxe2x80x21, Rxe2x80x22 and Rxe2x80x23 independently from each other represent xe2x80x94CH3 or xe2x80x94C2H5;
X is F, Cl, Br or I,
nxe2x80x2 is 1, 2, 3 or 4,
and wherein * denotes an asymmetric carbon and xe2x80x94 xe2x80x94 xe2x80x94 represents a single saturated bond or a double unsaturated bond.
Furthermore, the present invention concerns processes for the preparation of the above new compounds.
Thus, in one aspect, it is herein described a method to effect the impairment or destruction of a target biological substrate which method comprises treating the target substrate with an amount of the compound of formula I, Ixe2x80x2 or Ixe2x80x3 effective to photosensitize said substrate followed by irradiating said target substrate with radiation in a wavelength band absorbed by the compound of formula I, Ixe2x80x2 or Ixe2x80x3 for a time effective to impair or destroy the substrate.
In other aspect, the invention is therefore directed to pharmaceutical compositions comprising at least a compound of formula I, Ixe2x80x2 or Ixe2x80x3 as an active agent, together with a pharmaceutically acceptable carrier. The compositions are useful for in vivo photodynamic therapy and diagnosis of tumors and for killing of cells, viruses and bacteria, parasites and fungi in samples and in living tissues by well known photodynamic techniques.
Furthermore, the invention concerns the use of the compounds of formula I, Ixe2x80x2 or Ixe2x80x3 for the preparation of a pharmaceutical composition useful in photodynamic therapy.
The invention further concerns the use of the invention compounds for the preparation of compositions useful in diagnosis and ex vivo killing of bacteria, parasites, viruses and fungi.
The invention further concerns the acid chloride and anhydride of formulas II and III herebelow, respectively, as intermediates. 